Efecto de la adición de aceite esencial de canela americana ( Ocotea quixos ) en la permeabilidad al vapor de agua de películas de quitosana J. Food Sci. Gastron . (January - June 2024) 2 (1): 6-13https://doi.org/10.5281/zenodo.13996191ISSN: 3073-1283 ORIGINAL ARTICLE Effect of the addition of American cinnamon essential oil ( Ocotea quixos ) on the water vapor permeability of chitosan films Flor M. Fon-Fay ffonfay@uteq.edu.ec1 Instituto de Farmacia y Alimentos, Universidad de La Habana, Cuba.2 Facultad de Ciencias de la Ingeniería, Universidad Técnica Estatal de Quevedo, Los Ríos, Ecuador.Received: 23 May 2023 / Accepted: 27 September 2023 / Published online: 26 January 2024© The Author(s) 2024 Inalvis Escalante 1 · Flor M. Fon-Fay 2 · Jorge A. Pino 13 Abstract The objective of this study was to develop chi- tosan flms with Tween 80 and the essential oil of American cinnamon ( Ocotea quixos ) with good water vapor barrier properties. The thickness of the flms (42-92 µm) was consis - tent with those reported for similar biomaterials. The mois - ture content ranged between 23 and 48%, showing no sig - nifcant trend ( p >0.05) about the concentrations of chitosan and essential oil, likely due to the low amounts of essential oil added (0.1; 0.3; 0.5% v/v). The water vapor permeability (WVP) values ranged from 0.349 to 0.802 g mm m -2 h -1 kPa -1 , with no relevant changes due to polymer concentration or the addition of essential oil. A cubic model explained 99.76% of the variability in WVP, with a confdence level of 95%. The optimal formulation was 1.5% (m/v) chitosan, 0.3% (v/v) Tween 80, and 0.5% (v/v) essential oil. The optimized flm exhibited consistent properties in WVP, thickness, and moisture content with the other formulations, due to the stan - dardization of the flm production process. The addition of essential oil reduced the water solubility of the flms. Keywords chitosan flms, American cinnamon essential oil, barrier properties, water vapor permeability. Resumen El objetivo de este trabajo fue desarrollar pelícu - las de quitosana con Tween 80 y aceite esencial de canela americana ( Ocotea quixos ) con buenas propiedades de bar - rera al vapor de agua. Los espesores de las películas (42-92 µm) fueron consistentes con los reportados para biomateri - ales similares. El contenido de humedad osciló entre 23 y 48 %, sin mostrar una tendencia signifcativa ( p >0,05) en función de las concentraciones de quitosana y aceite esen - cial, probablemente debido a las bajas cantidades de aceite esencial añadidas (0,1; 0,3; 0,5 % v/v). Las permeabilidades al vapor de agua (WVP) variaron entre 0,349 y 0,802 g mm m -2 h -1 kPa -1 , sin cambios relevantes por la concentración de polímero o la adición del aceite esencial. Un modelo cúbico explicó el 99,76 % de la variabilidad de la WVP con un nivel de confanza del 95 %. La formulación óptima fue de 1,5 % (m/v) de quitosana, 0,3 % (v/v) de Tween 80 y 0,5 % (v/v) de aceite esencial. La película optimizada mostró propiedades consistentes en WVP, espesor y contenido de humedad con las demás formulaciones, debido a la estandarización del proceso. La adición de aceite esencial redujo la solubilidad de las películas en agua. Palabras clave películas de quitosana, aceite esencial de canela americana, propiedades de barrera, permeabilidad al vapor de agua. How to cite Escalante, I., Fon-Fay, F.M., & Pino, J.A. (2024). Efect of the addition of American cinnamon essential oil (Ocotea quixos) on the water vapor permeability of chitosan flms. Journal of Food Science and Gastronomy , 2 (1), 6-13. https://doi.org/10.5281/zenodo.13996191 3 Instituto de Investigaciones para la Industria Alimentaria, La Habana, Cuba.
J. Food Sci. Gastron . (January - June 2024) 2 (1): 6-13 7 Introduction In the past, consumers demanded more natural foods that were organoleptically and nutritionally similar to fresh products, while also being safe, hygienic, and having a long shelf life. Meeting these growing demands drove signifcant technological advancements in food packaging (Fadiji & Pathare, 2023).Active packaging techniques emerged as one of the most interesting innovations in this context, allowing for favor - able interactions between the packaging and the product to enhance quality and acceptability (Salgado et al., 2021). An example of active packaging for food includes edible flms and coatings that can incorporate active substances into their formulations. Their use in the food industry generated con - siderable interest due to their potential to extend the shelf life of many food products (Priya et al., 2023). These flms and coatings are made from biopolymers such as polysaccha - rides, proteins, and lipids, which are edible and biodegrad - able, making them non-toxic to the environment (Hashemi et al., 2023).Among the polysaccharides used in the preparation of ed - ible flms and coatings is chitosan, the main derivative of chitin, which is obtained industrially through chemical or en - zymatic deacetylation. The primary source of chitin comes from the exoskeletons of industrially processed crustaceans such as lobster, crab, and shrimp, contributing to the utiliza - tion of these waste products and reducing their environmen - tal impact (Ngasotter et al., 2023).Chitosan is noted for its excellent flm-forming and me - chanical properties, with no limitations regarding biocom - patibility, biodegradability, and toxicity. Additionally, it is naturally abundant and renewable (de Sousa et al., 2020). The antimicrobial properties of chitosan solutions and flms have been reported in several studies, demonstrating this biopolymer’s ability to inhibit the growth of a wide variety of bacteria (Khubiev et al., 2023). Although its antioxidant properties are limited, research has focused on incorporating antioxidant and antimicrobial substances into chitosan flms to enhance these characteristics.Essential oils have been included in chitosan polymer matrices due to their efective antimicrobial and antioxidant efects (Casalini & Giacinti, 2023). As natural compounds, they are biodegradable, leaving no residues, and do not harm the environment (Ponnusamy & Mani, 2022). The essential oil from the leaves of Ishpink ( Ocotea quixos ) has a high capacity to inhibit the growth of strains such as Staphylo-coccus epidermidis , Staphylococcus aureus , Escherichia coli , Streptococcus pyogenes , Streptococcus mutans , and the yeast Candida albicans (Valarezo et al., 2021). These char - acteristics give the essential oil the ability to enhance the bi - ological properties of chitosan flms when incorporated as an active substance, without afecting the inherent characteris - tics of the flm. Therefore, the following general objective was proposed: to evaluate the infuence of the concentrations of polymer, Tween 80, and essential oil of American cinna - mon ( O. quixos ) on the thickness and moisture content of chitosan flms. Materials and methods The research was conducted in the laboratories of the In - stitute of Pharmacy and Food at the University of Havana, as part of the project on the extraction of chitosan and its salts from lobster chitin ( Panulirus argus ) for pharmaceutical and food applications. The experimental design and analysis of water vapor permeability (WVP) of chitosan flms with Tween 80 and essential oil of American cinnamon ( O. quixos ) were per - formed using Design Expert 8.0.6 software (Stat-Ease Inc., Minneapolis, USA). The flm with the lowest WVP value was selected using a cubic response surface model for nu - merical optimization. The evaluated factors included the concentration of essential oil (A), chitosan (B), and Tween 80 (C), while WVP served as the response variable. The soft - ware defned 18 experimental combinations, including three replicates (Table 1). Table 1. Experimental design matrix RunEssential oil (% v/v)Chitosan (% m/v)Tween 80 (% v/v) 10.52.00.520.11.50.130.52.00.340.51.50.150.51.50.560.12.00.170.12.00.380.11.50.590.32.00.1100.32.00.5110.52.00.1120.31.50.3130.31.50.5140.32.00.5150.31.50.3160.11.50.1170.12.00.5180.32.00.3Chitosan (221 kDa and 76% degree of deacetylation) was used, obtained through thermo-alkaline N-deacetylation of
J. Food Sci. Gastron . (January - June 2024) 2 (1): 6-13 8 chitin from common lobster ( P. argus ) at the Production Plant for Natural and Synthetic Products of the Center for Research and Development of Medicines (Havana, Cuba) (de la Paz et al., 2012). Other materials used included the essential oil of American cinnamon ( O. quixos ) supplied by the Chankuap Foundation (Ecuador), Tween 80 (Acros Or - ganics, Belgium), 90% lactic acid (Merck, Germany), and distilled water.The flm-forming solutions were prepared with chitosan at 1.5% and 2.0% (m/v) in a 1% lactic acid solution and agitated for 2 hours. Tween 80 was then added at various concentra - tions according to the experimental design, and the mixture was fltered to remove impurities. Next, the essential oil of cinnamon was incorporated, and the mixture was emulsifed at 12000 min⁻¹ for 5 minutes using an Ultra-Turrax homoge - nizer. The emulsions were allowed to rest for deaeration and poured into glass molds, drying at 40 °C for 24 hours.The concentrations of chitosan were selected based on pre - vious studies on the physical properties of the flms (Casar - iego, 2009). The obtained flms were stored in double-sealed Ziploc® bags within a desiccator, maintained at a controlled relative humidity of 21-22%.To evaluate the physical properties of the chitosan flms, thickness, moisture content, water solubility, and water va - por permeability (WVP) were measured. The thickness of the flms was determined using a digital micrometer with an accuracy of ±1 µm. Moisture content was evaluated by weighing the samples before and after drying at 105 ºC for 24 hours, while water solubility was measured by immers - ing the flms in distilled water at room temperature for 24 hours and calculating the percentage of dissolved material. WVP was determined following the standard gravimetric method using permeable capsules and measuring weight loss through the flms at 25 ºC and relative humidity of 50%. The results obtained were statistically analyzed using Statistica software. Results and discussion Table 2 shows the thickness and moisture content values of chitosan flms with the essential oil of American cinnamon ( O. quixos ). The thickness of the flms ranged from 42 to 92 µm, which could be attributed to diferences in moisture content, as protonated chitosan has a high afnity for water, increasing its hydrophilicity compared to its powdered form (Desai et al., 2023). During flm formation, the interaction between chitosan and lactic acid increases water absorption, likely due to the loss of integrity in the molecular structure, which exposes more functional groups for water absorption (Yadav et al., 2023). Table 2. Efect of the addition of essential oil of American cinnamon ( O. quixos ) on the thickness and moisture content of chitosan flms RunEssential oil (% v/v)Chitosan (% m/v)Tween 80 (% v/v)Thickness (µm)Moisture (% m/m) 10.52.00.572 (2) abcd32 (5) bcdefg20.11.50.173 (9) abcd31 (8) cdefgh30.52.00.368 (6) bcd38 (4) bc40.51.50.192 (5) a29 (4) defgh50.51.50.568 (1) bcd25 (4) gh60.12.00.161 (9) bcde25 (3) gh70.12.00.357 (7) cde28 (1) efgh80.11.50.575 (1) abcd27 (6) fgh90.32.00.158 (2) bcde23 (3) h100.32.00.553 (7) de32 (3) bcdefg110.52.00.142 (5) e35 (3) bcdef120.31.50.365 (4) bcd35 (1) bcdef130.31.50.559 (1) bcde36 (5) bcde140.32.00.561 (8) bcde37 (4) bcd150.31.50.379 (2) ab48 (3) a160.11.50.177 (1) abc36 (4) bcdef170.12.00.558 (4) bcde40 (1) b180.32.00.378 (9) abc32 (3) bcdefg Mean (Standard deviation).Diferent letters indicate signifcant diferences ( p ≤0.05).
J. Food Sci. Gastron . (January - June 2024) 2 (1): 6-13 9 The thickness values obtained in this study are generally lower than those reported by Peng et al. (2013), who report - ed thicknesses ranging from 72 to 131 µm for 2% (m/v) chi - tosan flms with green and black tea extracts, acetic acid, and glycerol. Conversely, Moradi et al. (2012) reported thickness values ranging from 70 to 80 µm for 2% chitosan flms in acetic acid with essential oil of Zataria multifora Boiss and grape seed extract, using glycerol as a plasticizer. Under sim - ilar conditions but with the addition of Tween 80, Ojagh et al. (2010) obtained values between 95 and 107 µm.In general, the thickness values obtained in this research fall within the range reported in the literature. However, the thickness of the flms can vary depending on factors such as temperature and humidity, which are difcult to control, leading many authors not to report this parameter in studies of biodegradable flms.Table 2 shows that the moisture content of the flms var - ied between 23 and 48% (m/m), with no signifcant trend ( p >0.05) observed concerning the concentrations of chitosan and essential oil, which could be attributed to the low con - centrations of essential oil used (0.1, 0.3, and 0.5% v/v). Ojagh et al. (2010) reported a decrease in moisture content with increasing concentrations of essential oils (0.4 to 2%) in 1, 2, and 3% chitosan flms in acetic acid. However, Bo - nilla et al. (2013) indicated that the addition of hydrophobic compounds to 1% chitosan flms did not signifcantly afect ( p >0.05) moisture content, likely due to the low solid con - tent. In contrast to the results of this study, Peng et al. (2013) observed a decrease in moisture content in 2% chitosan flms with glycerol and green and black tea extracts (0.5%), re - ducing from 28 to 19% and 23%, respectively. Wang et al. (2013) reported similar results with the addition of tea poly - phenols (10-40%), with values ranging from 42 to 25%.A key function of flms is to prevent the transfer of mois - ture between the food and its environment (Azevedo et al., 2022). Water vapor permeability (WVP) develops in three phases: absorption of water from the area of higher relative humidity, difusion through the flm, and desorption in the area of lower relative humidity (Turan et al., 2021). The WVP values for chitosan flms with essential oil of Ameri - can cinnamon were calculated from the WVTR results (with R² between 0.9989 and 0.9999) and are presented in Table 3. Neither the concentration of chitosan nor the addition of essential oil signifcantly modifed the WVP, with values ranging from 0.349 to 0.802 g mm m -2 h -1 kPa -1 . Bonilla et al. (2013) and Pranoto et al. (2005) also found that the addition of basil, thyme extracts, and garlic essential oil did not nota - bly afect this property. Table 3. Efect of the addition of essential oil of American cinnamon ( O. quixos ) on the water vapor permeability of chi - tosan flms RunEssential oil (% v/v)Chitosan (% m/v)Tween 80 (% v/v)R 2 WVTR(g/h m 2 )WVP(g mm/m 2 h kPa) 10.52.00.50.998937.1 (0.6)0.566 (0.009) cdef20.11.50.10.999436.2 (3.1)0.505 (0.03) cdefg30.52.00.30.998935.4 (0.7)0.479 (0.06) efg40.51.50.10.999333.5 (1.9)0.588 (0.1) bcde50.51.50.50.999733.8 (0.7)0.469 (0.003) efgh60.12.00.10.999734.4 (2.9)0.456 (0.07) fghi70.12.00.30.999633.7 (1.0)0.602 (0.05) bcd80.11.50.50.999932.5 (0.5)0.608 (0.03) bc90.32.00.10.999634.9 (1.8)0.697 (0.03) b100.32.00.50.999933.4 (1.1)0.526 (0.07) cdefg110.52.00.10.999836.7 (0.3)0.486 (0.1) defg120.31.50.30.999731.8 (0.6)0.363 (0.04) hi130.31.50.50.999631.8 (0.5)0.544 (0.05) cdef140.32.00.50.999732.5 (1.8)0.546 (0.02) cdef150.31.50.30.999634.6 (3.8)0.349 (0.03) i160.11.50.10.999132.3 (1.1)0.525 (0.008) cdefg170.12.00.50.998932.7 (0.8)0.802 (0.01) a180.32.00.30.999231.7 (1.2)0.414 (0.1) ghi WVRT: water vapor transmission rate; WVP: water vapor permeability. Mean (Standard deviation). Diferent letters indicate signifcant diferences ( p ≤0.05).
J. Food Sci. Gastron . (January - June 2024) 2 (1): 6-13 10 Several researchers (Peng et al., 2013; Wang et al., 2013; Siripatrawan and Harte, 2010) have demonstrated that the addition of polyphenols reduces WVP. In the study by Peng et al. (2013), a decrease of up to 62.1% compared to the con - trol was observed when using a 2% extract. Siripatrawan & Harte (2010) attribute this decrease to the interaction be - tween polyphenols and the chitosan structure.Although the general patterns observed in this study and previous ones do not coincide, the WVP values with essen - tial oil concentrations and extracts similar to those in our re - search are comparable. The diferences could be related to the fact that all these flms were made with 2% chitosan in an acetic acid solution and glycerol as a plasticizer. Acetic acid has been shown to produce flms with lower WVP compared to lactic acid, while the addition of a plasticizer tends to in - crease WVP (Eslami et al., 2023). Furthermore, factors such as the type and amount of plasticizer and solvent, as well as the molecular weight and degree of deacetylation of chi - tosan, may also infuence permeability (Wang et al., 2013). The infuence of active compounds on reducing WVP was reported by Ojagh et al. (2010) when evaluating the perme - ability of flms with cinnamon essential oil, fndings that co - incide with those of Pastor et al. (2013) on the incorporation of resveratrol. On the other hand, Bonilla et al. (2013) found that citric acid and α-tocopherol had no signifcant impact ( p >0.05) on WVP values.The results of the analysis of variance for regression and the estimated coefcients related to the water vapor perme - ability of chitosan flms. The best ft was achieved with a cubic model, which was signifcant at a 95.0% confdence level, indicating a signifcant relationship between the fac - tors and the dependent variable of the model. The R² statistic showed that the ftted model explains 99.76% of the variabil - ity in WVP. The cubic term of the essential oil concentration and the linear interaction of the three factors did not signif - cantly afect the WVP of the flms.Figure 1 illustrates the infuence of these factors on the WVP of chitosan flms with Tween 80 and the essential oil of American cinnamon. It is observed that the addition of Tween 80 at 0.3% (v/v) improved the water vapor barrier properties of chitosan flms with essential oil of American cinnamon ( O. quixos ), likely due to the hydrogen bonds es - tablished between the polar groups of chitosan and the polar groups of Tween 80, thereby reducing the number of polar groups available to interact with water molecules (Bide et al., 2021). Figure 1. Water vapor permeability of chitosan flms with essential oil of American cinnamon ( O. quixos ) and Tween 80 at concentrations of a) 0.1; b) 0.3; and c) 0.5% (v/v). --- Chitosan 1.5% --- Chitosan 2% (m/v).The mechanism predicting water transport through hydro - philic flms, such as those made of chitosan, is complex due to the non-linear nature of the absorption isotherms and the fact that difusivity varies with water content (Souza et al., 2009). Additionally, the water vapor fow through these flms behaves non-linearly about the partial vapor pressure gra - dient. If the flms are cationic and highly hydrophilic, they
J. Food Sci. Gastron . (January - June 2024) 2 (1): 6-13 11 may interact with the polymer matrix, which could increase the WVP.Villalobos et al. (2006) found that the WVP of hydroxypro - pyl methylcellulose flms with a mixture of Span 60 and su - crose esters, which had HLB values between 4.7 and 8.0, was lower than that of control flms; moreover, WVP decreased with low ratios of hydrocolloid to surfactant. Rodríguez et al. (2006) reported that the WVP of starch flms was not sig - nifcantly afected by the addition of Tween 20 and lecithin at concentrations of 0.5 to 5%. On the other hand, Chen et al. (2009) observed that the WVP of starch and decolorized hsian-tsao gum flms with surfactants (sucrose esters S-0770 with HLB = 7, S-1170 with HLB = 11, and S-1570 with HLB = 15) was signifcantly lower compared to control flms, and this decrease was correlated with the increase in HLB values of the surfactants, indicating that the efect of the surfactant on WVP depends on its type and concentration, as well as the properties of the flm-forming material. The verifcation of the normality assumption was con - ducted through an analysis of variance, analyzing the nor - mal probability of the residuals. The values of the internally studentized residuals ft a straight line, indicating a normal distribution of errors and confrming the normality hypoth - esis. For the numerical optimization of the formulation of chitosan flms with essential oil of American cinnamon, the ranges of the independent variables (concentrations of chi - tosan, Tween 80, and essential oil) were used as constraints to achieve the lowest WVP values in the flms (Table 4). Table 4. Constraints for the optimization of the formulation of chitosan flms with the essential oil of American cinnamon ( Ocotea quixos ) ParameterLower limitUpper limitCriterion Essential oil (% v/v)0.10.5In the rangeChitosan (% m/v)1.52.0In the rangeTween 80 (% v/v)0.10.5In the rangeWVP (g mm/m 2 h kPa)0.3491960.8023MinimizeThe program suggested 25 optimized solutions for the for - mulation of chitosan flms with the essential oil of American cinnamon based on the previous constraints. Solution 5 was selected, as it had the lowest WVP and high statistical con - venience, corresponding to the lowest percentage of chitosan and the highest of essential oil.Table 5 shows the results of the evaluated properties of the flm obtained from the optimized formulation. The WVP of the optimized flm was higher than the value estimated from the numerical optimization of the flm formulation, although the diferences are practically insignifcant. Table 5. Properties of the optimized chitosan flm with Tween 80 and the essential oil of American cinnamon (n = 3) Parameter Mean (Standard deviation) Water vapor permeability (g mm/m 2 h kPa)0.50 (0.02)Thickness (µm)64 (1)Solubility (% m/m)34 (2)Moisture (% m/v)35 (4) WVP: Water vapor permeability. The incorporation of essential oil of American cinnamon into chitosan flms resulted in a decrease in their water sol - ubility. Rodríguez (2015) reported solubility values of 49 and 47% for chitosan flms at 1.5 and 2%, respectively, with thicknesses similar to those in this study. Ojagh et al. (2010) documented a water solubility of 23.2% for chitosan flms that were 95 ± 2.5 μm thick at 2% (w/v) in a 1% (v/v) acetic acid solution, which was lower than the results obtained in this work, supporting the hypothesis that greater thickness results in lower water solubility. Wang et al. (2013) reported a solubility of 23.5% for flms at 4% (w/v) in 2% (v/v) acetic acid, showing a similar trend, although without signifcant diferences. Despite the concentration of essential oil used to reduce solubility, Ojagh et al. (2010) observed a signifcant reduction in this indicator starting at 1.5% of the essential oil of cinnamon. On the other hand, Peng et al. (2013) reported an increase in solubility upon adding tea extracts. Conclusions Chitosan flms with the essential oil of American cinna - mon ( O. quixos ) exhibited thicknesses consistent with those reported for this type of biomaterial. The moisture content in the flms did not show a signifcant trend concerning the concentrations of chitosan and essential oil, which could
J. Food Sci. Gastron . (January - June 2024) 2 (1): 6-13 12 be attributed to the low amounts of essential oil used in the formulation. Additionally, water vapor permeability was not afected by either polymer concentration or the addition of essential oil. A cubic model was found to be the most suit - able for explaining the variability in water vapor permeabil - ity, resulting signifcantly in a 95.0% confdence level. The optimal formulation for the flms was identifed and related to permeability properties. The permeability, thickness, and moisture content characteristics of the optimized flm were consistent with those of the other analyzed flms, indicating standardization in the laboratory-scale production process. Furthermore, the incorporation of the essential oil of Amer - ican cinnamon reduced the solubility of the flms in water. 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